Obtaining And Progeny Analysis Of Transgenic Glyphosate-resistant Soybean

Soybean（Glycine max Merr.（L.））is an important food crop in China.With the development of modern biotechnology,transgenic technology has become one of the important means of genetic improvement of soybean.In this study,glyphosate-resistant gene G10-EPSPS was transferred into receptor soybeans through agrobacterium-mediated transformation of cotyledonary nodes.Then new herbicide-resistant soybean varieties with independent intellectual property rights would be cultivated in China.And transgenic glyphosate-resistant soybean（cultivated by our own laboratory）SHZD32-01 was used as material to evaluate its herbicide-tolerance and survival competitiveness in cultivated land.This study aims to provide materials and basic data for industrialization of transgenic herbicide-resistant soybean in China.The results are as follows.1.Dongnong 50 and Wandou 28 were used as genetic transformation receptors.Two Dongnong 50 regeneration plants and one Wandou 28regeneration plants（W28）were obtained.The results of PCR and glyphosate spraying on leaves of three plants were positive in T₀generation.When two strains of Dongnong 50 transformants were planting in artificial climate chamber,their leaves became dark green and waxy,and couldn’t bear normally.W28 in T₀ generation growed quickly and healthily.It has a good glyphosate resistance.2.W28 in T₀ generation was heterozygous.Its glyphosate resistance separated in T₁ generation.The RT-PCR and RT-qPCR results showed that G10-EPSPS gene had very high expression in the root,stem and leaf of T₁positive plants.And the gene was expressed to the highest level in leaves,the second level in stem,flower and young pod.The expression amount in stem,flower and young pod were equally likely.The gene had the lowest expression in root.Western blot results showed EPSPS protein was expressed in all organs,including root,stem,leaf,flower,and fruit.3.The growth of transgenic glyphosate-resistant soybean SHZD32-01was not inhibited after glyphosate treatment at 1230-4920 g a.i./hm².In cultivated land,there were no significant differences of the growth indices of SHZD32-01 and control varieties.The indices included plant height,compound leaf number,coverage degree and propagation coefficient,shattering rate.Therefore SHZD32-01 had no competitive advantage and low weediness potential.